Investigating the Time-Dependent Reversibility of DCM-Induced Epigenetic, Matrix, and Functional Remodeling

研究 DCM 诱导的表观遗传、基质和功能重塑的时间依赖性可逆性

• 批准号: 10535260
• 负责人: Isabella Reichardt
• 金额: $ 4.16万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-16 至 2025-09-15
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10535260
• 关键词: ATAC-seq; Aftercare; Age; Age-Months; Cardiac; Cardiac Myocytes; Cardiology; Cardiovascular Diseases; Cardiovascular system; Cells; Chromatin; Collagen; Congestive Heart Failure; Coupled; Cytoskeleton; DNA Methylation; Data; Deposition; Dilated Cardiomyopathy; Disease; Dose; Doxycycline; EFRAC; Engineering; Ensure; Epigenetic Process; Etiology; Extracellular Matrix; Family suidae; Fellowship; Fibroblasts; Fibrosis; Functional disorder; Future; Gene Expression; Generations; Genes; Genetic Transcription; Genome; Genomics; Goals; Heart; Heart Diseases; Heart Hypertrophy; Heart failure; Human; Hypertrophy; In Vitro; Induced Mutation; Inherited; Injury; Learning; Left ventricular structure; Link; Mechanics; Medicine; Memory; Mentors; Mentorship; Mesenchymal Stem Cells; Microfilaments; Modeling; Mus; Muscle Cells; Mutate; Mutation; Myocardial dysfunction; Myocardium; Myosin ATPase; Nature; Organ; Outcome; Patients; Periodicity; Pharmacological Treatment; Pharmacology; Phenotype; Physicians; Plant Roots; Point Mutation; Progressive Disease; Proteomics; Research; Resolution; Running; Scientist; Signal Transduction; Structure; Symptoms; Technology; Testing; Therapeutic; Time; Training; Transgenic Mice; Treatment Failure; Troponin C; United States National Institutes of Health; career; career development; chromatin remodeling; clinically relevant; effective therapy; epigenetic memory; epigenome; experimental study; familial dilated cardiomyopathy; healing; heart function; improved; inhibitor; mechanical stimulus; mouse model; multiple omics; mutant; palliative; pre-doctoral; pressure; professor; response; screening; therapeutic target; tool; transcriptome sequencing; transgene expression; translational therapeutics

Project Summary Dilated cardiomyopathy (DCM) is a highly prevalent inherited cardiac disease, characterized by systolic dysfunction, eccentric hypertrophy, and left ventricle dilation. While pharmacologic and mechanical treatments have been shown to partially improve cardiac function, these results are often short lived and highly variable from patient to patient. Moreover, recent studies have demonstrated that epigenetic and matrix dysregulation can persist, even in patients with improved systolic function after treatment. Given that aberrant chromatin remodeling and extracellular matrix (ECM) deposition have been identified as drivers of dilated remodeling—and that chromatin and ECM remodeling can become irreversible over time—it is crucial to understand the time- dependent effects of DCM mutations on reversing maladaptive remodeling at the genome, myocyte, and matrix levels. The central hypothesis of this proposal is that the reversibility of the DCM phenotype is time-dependent due to the accumulation of permanent ECM and chromatin remodeling as the disease progresses. To complete this proposal, I will utilize a DCM mouse model created by the Davis lab which contains a mutation (I61Q) in cardiac troponin C (cTnC) that directly decreases the myofilament’s Ca2+ sensitivity, leading to eccentric hypertrophy, systolic dysfunction, and left ventricle dilation. Importantly, expression of this mutant can be temporally controlled with doxycycline to specifically test our central hypothesis. In this proposal I will 1) Determine the time-dependent effects of I61Q cTnC expression on myocyte, matrix, and chromatin accessibility, 2) Examine the reversibility of DCM remodeling in myocyte, matrix, and chromatin accessibility at different stages of the disease, and 3) Determine if myocytes retain epigenetic memories of the mechanical disequilibrium caused by DCM. Improving our understanding of the time-dependent reversibility of DCM remodeling will better inform therapeutic targets and treatment windows for patients with DCM. Moreover, completion of this project will enhance Bella’s training as an independent scientist and prepare her to one day become a professor in cardiovascular engineering. Receiving the NIH F31 predoctoral fellowship will facilitate important experiments and training that will aid in her pursuit of this career goal. In this project, Bella will gain expertise in multi-omic approaches—such as proteomics, RNAseq, and ATACseq—which are growing in popularity due to their unbiased screening capabilities. The UW Genomics Core will assist Bella in learning how to effectively use these tools, which will not only benefit this project but will also be an incredibly useful skillset for Bella’s future career. Given the clinical relevance of this project, we have engaged Farid Moussavi-Harami, MD, an acting physician- scientist who practices cardiology within the UW Department of Medicine. Dr. Moussavi-Harami’s input as a clinician will be crucial for ensuring our research questions and aims remain relevant to patients, and his mentorship throughout this project will enhance Bella’s training and career development as she aims to eventually run a lab with an emphasis in translational therapeutics and technologies for cardiovascular diseases.

项目摘要 扩张型心肌病(DCM)是一种高度流行的遗传性心脏病，其特点是收缩 心功能不全、偏心性肥厚和左心室扩大。虽然药物治疗和机械治疗 已经被证明可以部分改善心脏功能，但这些结果往往是短暂的和高度可变的 从一个病人到另一个病人。此外，最近的研究表明，表观遗传和基质失调 可以持续，即使在治疗后收缩功能改善的患者。鉴于这种异常的染色质 重构和细胞外基质(ECM)沉积已被确定为扩张性重构的驱动因素--和 染色质和细胞外基质的重塑可能会随着时间的推移而变得不可逆转-了解时间是至关重要的- DCM突变对逆转基因组、心肌细胞和基质不良适应性重构的依赖作用 级别。这一建议的中心假设是DCM表型的可逆性是时间相关的 由于随着疾病的进展，永久性的细胞外基质和染色质重塑积聚。要完成 在这项建议中，我将利用Davis实验室创建的DCM小鼠模型，该模型包含一个突变(I61Q) 心肌肌钙蛋白C(CTNC)直接降低肌丝对钙的敏感性，导致离心性 肥厚、收缩功能障碍和左心室扩张。重要是，该突变体的表达可以是 用多西环素临时控制，以具体测试我们的中心假说。在此提案中，我将1) 确定I61Q cTNC表达对心肌细胞、基质和染色质可及性的时间依赖性影响， 2)检测心肌细胞、基质和染色质在不同阶段的可逆性。 以及3)确定心肌细胞是否保留了机械失衡引起的表观遗传记忆 由DCM提供。提高我们对DCM重塑的时间依赖性可逆性的理解将更好地了解 扩张型心肌病患者的治疗靶点和治疗窗口。此外，该项目的完成将 加强贝拉作为独立科学家的培训，并为她有朝一日成为一名 心血管工程学。获得NIH F31博士前奖学金将促进重要的实验 和训练，这将有助于她追求这个职业目标。在这个项目中，贝拉将获得多组学方面的专业知识 蛋白质组学、RNAseq和ATACseq等方法由于其 无偏见的筛查能力。威斯康星大学基因组学核心将帮助贝拉学习如何有效地使用这些 工具，这不仅将使这个项目受益，也将是贝拉未来职业生涯中非常有用的技能集。 鉴于这个项目的临床相关性，我们聘请了代理内科医生法里德·穆萨维-哈拉米医学博士- 在威斯康星州医学部从事心脏病学工作的科学家。穆萨维-哈拉米博士作为 临床医生将是确保我们的研究问题和目标与患者相关的关键，而他的 整个项目的指导将加强贝拉的培训和职业发展，因为她的目标是 最终建立一个实验室，重点研究心血管疾病的转化疗法和技术。